ANALYSIS OF BACTERIAL CONSTITUENTS OF THE ACTIVE BENIGN CHRONIC SUPPURATIVE
OTITIS MEDIA WITH RIBOSOMAL INTERGENIC SPACER ANALYSIS-POLYMERASE CHAIN
REACTION APPROACH
Novi Primadewi, Soepomo Soekardono, Sri Winarti , Anton Christanto
Department of Otolaryngology Head and Neck Surgery, Faculty of Medicine,
University of Gadjah Mada/DR. Sarjito Hospital, Yogyakarta 55281
ABSTRACT
Background: Active benign chronic suppurative otitis media (ABCSOM) is a
potentially serious disorder, since there is a risk of permanent hearing lost,
and mainly caused by bacterial infections. Traditionally, the identification
and enumeration of bacterial species has depended entirely upon pure-culture
techniques. However, the difficulty with which some types of organisms are
cultured, particularly fastidious and/or anaerobic bacteria means that the more
easily grown species in a mixed microbial community are likely to be
overrepresented by cultivation and plate counts. Bacterial molecular-analyses
have provided a new perspective on bacterial analysis by allowing the detection
and identification of constituent microorganisms in the absence of cultivation.
Objective: In this study we established a culture-independent by using
ribosomal intergenic spacer analysis (RISA) to survey bacteria present in the
active benign chronic suppurative otitis media without fastidious cultivation.
Methods: Middle ear fluid discharge samples were collected from 5 individuals
of ABCSOM using sterile syringe equipped with intravenous catheter. Samples
collected in the laboratory were preserved with isopropanol and immediately
frozen and stored at -20°C prior to DNA extraction. Total DNA was direct
extracted from the samples, and the bacterial constituents were analyzed by
RISA.
Results: Several bacterial species were detected using RISA, whereas culture
dependent examination based approach (plate method) revealed only one type of
bacterium from all samples.
Conclusion: The results showed that culture-independent approach (RISA) is the
robust method to determine bacterial constituent middle ear fluid discharge.
Further investigation such sequencing analysis is needed to determine
phylogenetic affiliation of bacterial species residing in middle ear fluid
discharge.
Key words: ABCSOM, ribosomal intergenic spacer analysis, culture independent,
bacterial community.
dr. HM Anton Christanto
Bag/SMF THT RSUP Dr Sardjito-FK UGM
Jl. Kesehatan no 1 Yogyakarta - Indonesia 55284
Phone/Fax : +62 274 7474230
Mobile +62(0)8122658495
Email : [EMAIL PROTECTED]
[EMAIL PROTECTED]
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